We will continue to investigate at both cellular and molecular levels, the mechanisms by which adeno-associated satellite viruses (AAV) interfere with the replication and potential oncogenesis of their helper adenoviruses and herpesviruses. We will use conditionally defective helper viruses, namely temperature-sensitive mutants of human adenovirus type 31 and herpes simplex virus, type 1, known to be defective in viral DNA synthesis and/or other virus-specific functions. Complementation studies with these mutants will simplify dissection of the defective satellite replication cycle. AAV are defective at both the replication and maturation steps. We will analyze the products synthesized in these complex systems by the techniques of radioisotope labeling of viral DNA, ultracentrifugal analysis, electron microscopy, column chromatography and polyacrylamide gel electrophoresis, and correlate these parameters with biological function. We will initiate experiments to clone AAV DNA in E. coli in an effort to obtain relatively large quantities of this reagent by a simpler and more economical procedure than that currently available.